Use of pridopidine to improve cognitive function and for treating alzheimer&#39;s disease

ABSTRACT

This invention provides a method of improving cognitive function in a subject comprising periodically administering to the subject an amount of pridopidine or a pharmaceutically acceptable salt thereof effective to improve a cognitive function in the subject. 
     The invention also provides a method of treating a subject afflicted with Alzheimer&#39;s disease, comprising periodically administering to the subject a pharmaceutical composition comprising an amount of pridopidine or a pharmaceutically acceptable salt thereof effective to treat the subject.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation-in-part application of U.S. patentapplication Ser. No. 15/052,368 filed Feb. 24, 2016, which claims thebenefit of U.S. Provisional Application No. 62/186,221, filed Jun. 29,2015, and U.S. Provisional Application No. 62/120,771, filed Feb. 25,2015, the entire contents of which are hereby incorporated by referenceherein.

Throughout this application, various publications are referred to byfirst author and year of publication. Full citations for thesepublications are presented in a References section immediately beforethe claims. Disclosures of the publications cited in the Referencessection are hereby incorporated by reference in their entireties intothis application in order to more fully describe the state of the art asof the date of the invention described herein.

BACKGROUND

Pridopidine (4-[3-(methylsulfonyl)phenyl]-1-propyl-piperidine, ACR16,Huntexil) is a drug under development from a new class of pharmaceuticalagents, the dopidines, which are considered to have dopaminergicstabilizing properties (U.S. Patent Publication No. US 2014/0378508).Dopaminergic stabilizers are compounds that can both enhance andcounteract dopamine dependent functions in the central nervous system(CNS), depending on the initial level of dopaminergic activity (U.S.Patent Publication No. US 2014/0378508). Dopaminergic stabilizerssuppress the hyperactive behavior induced by stimulants such asamphetamine. In contrast, at low levels of dopamine function, thedopamine stabilizers enhance behavioral activity (U.S. PatentPublication No. US 2014/0378508). The primary effect of pridopidine onHD-related motor symptoms is therefore expected to occur via thedopamine transmissions modulating properties of pridopidine (Ponten2010). Processes of synthesis of pridopidine and a pharmaceuticallyacceptable salt thereof are disclosed in U.S. Pat. No. 7,923,459. U.S.Pat. No. 6,903,120 claims pridopidine for the treatment of Parkinson'sdisease, dyskinesias, dystonias, Tourette's disease, iatrogenic andnon-iatrogenic psychoses and hallucinoses, mood and anxiety disorders,sleep disorder, autism spectrum disorder, ADHD, Huntington's disease,age-related cognitive impairment, and disorders related to alcohol abuseand narcotic substance abuse.

Alzheimer's Disease

Alzheimer's disease (AD) is the most common form of dementia, a generalterm for memory loss and other intellectual abilities serious enough tointerfere with daily life. AD accounts for 60 to 80 percent of dementiacases (www.alz.org).

AD is characterized by the loss of synapses and neurons from the brain,and by the accumulation of extracellular protein-containing deposits(referred to as ‘senile plaques’) and neurofibrillary tangles (Selkoe etal. 2001)

The most common early symptom of AD is difficulty remembering newlylearned information. As AD advances it leads to increasingly severesymptoms, including disorientation, mood and behavior changes, as wellas difficulty speaking, swallowing and walking.

Currently, there is no cure for AD. New effective therapies for AD areneeded.

DMN (Default Mode Network)

The default mode network (DMN), is a large-scale brain network ofinteracting brain regions known to have activity highly correlated witheach other and distinct from other networks in the brain. The DMNincludes brain regions with high degrees of functional connectivity andis active in the brain at rest but becomes deactivated when taskperformance is initiated. The clinical implications of the DMN inneurological and neuropsychiatric disorders have been, and continue tobe, targets of investigation. In disorders including AD, PD, TLE, ADHD,and mood disorders, the DMN has been implicated inneurobiological/neurocognitive pathophysiological models. (YALE JOURNALOF BIOLOGY AND MEDICINE 89 (2016), pp. 49-57).

DMN (Default Mode Network) plays crucial role in cognitive processing,and dysfunction in the DMN is associated with aberrant behaviors andcognition in various NDDs (F. Sambataro et al, 2010; F. Agosta et al,2012; Bonnelle V et al, 2012). The DMN is comprised of brain regionsknown to be involved in cognitive functions: posterior cingulate cortex(PCC), lateral parietal cortex and medial prefrontal cortex (mPFC), andprominent connections to nodes in the medial temporal lobe (MTL) andangular gyrus (Mevel K. et al, 2011; Buckner R. L. et al, 2008).

Specifically, aberrant DMN connectivity seems to be the most affectednetwork in Dementia due to AD (Balthazar et al, AAN 2014). In AD, theDMN consistently shows reduced connectivity, and has even been shown topredict disease severity and progression (Petrella J R, et al, 2011;Brier M R. et al, 2014).

Impaired DMN connectivity is also described in patients with FTD (ZhouJ, et al, 2010; Kipps C M. and Hodges J R., 2006), ALS (Girardi A, etal, 2011), PSP (Ghosh B C, et al, 2012), PD (Lucas-Jimenez et al, 2016),as well as in several neuropsychiatric diseases (Buckner R L et al,2008).

Additional number of studies identified DMN disturbances inschizophrenia, autism, hyperactivity disorder, epilepsy and multiplesclerosis (M. Guye et al, 2010).

SUMMARY OF THE INVENTION

This invention provides a method of improving cognitive function in ahuman subject comprising administering to the subject an amount ofpridopidine or a pharmaceutically acceptable salt thereof wherein thecognitive function in need for improvement is associated with DMNdysfunction.

This invention provides a method of treating a human subject afflictedwith a disease selected from Alzheimer's disease and mild cognitiveimpairment (MCI) comprising administering to the subject apharmaceutical composition comprising an amount of pridopidine or apharmaceutically acceptable salt thereof wherein said disease isassociated with DMN dysfunction.

In one embodiment, pridopidine increases the DMN functional connectivity(increased activity). In another embodiment, the increase in functionalconnectivity is in the insula. In another embodiment, the increase infunctional connectivity is in the parietal and temporal cortex. Inanother embodiment, the increase in functional connectivity is in theprecuneus.

This invention provides a method of treating a human subject afflictedwith epilepsy, comprising administering to the subject a pharmaceuticalcomposition comprising an amount of pridopidine or a pharmaceuticallyacceptable salt thereof.

This invention provides a method of improving cognitive function in asubject comprising periodically administering to the subject an amountof pridopidine or a pharmaceutically acceptable salt thereof effectiveto improve a cognitive function in the subject.

The invention also provides a package comprising:

a) a pharmaceutical composition comprising an amount of pridopidine or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier;

b) instructions for use of the pharmaceutical composition to treat asubject afflicted with a disease or disorder associated with a cognitivedeficit.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with a disease or disorderassociated with a cognitive deficit, which comprises:

a) one or more unit doses, each such unit dose comprising an amount ofpridopidine or a pharmaceutically acceptable salt thereof, wherein theamount of said pridopidine in said unit dose is effective, uponadministration to said subject, to treat the subject, and

b) a finished pharmaceutical container therefor, said containercontaining said unit dose or unit doses, said container furthercontaining or comprising labeling directing the use of said package inthe treatment of said subject.

The invention also provides a pharmaceutical composition comprising anamount of pridopidine or a pharmaceutically acceptable salt thereof foruse in treating a subject afflicted with a disease or disorderassociated with a cognitive deficit.

The invention also provides a pharmaceutical composition in a unitdosage form, useful in treating a subject afflicted with a disease ordisorder associated with a cognitive deficit, which comprises an amountof pridopidine or a pharmaceutically acceptable salt thereof, whereinthe amount of said pridopidine in said composition is effective, uponadministration to said subject of one or more of said unit dosage formsof said composition, to treat the subject.

The invention also provides a package comprising:

a) a pharmaceutical composition; and

b) instructions for use of the pharmaceutical composition to treat asubject afflicted with a disease or disorder associated with a cognitivedeficit.

This invention also provides pridopidine or a pharmaceuticallyacceptable salt thereof for use in treating a disease or disorderassociated with a cognitive deficit.

This invention also provides a use of pridopidine or a pharmaceuticallyacceptable salt thereof in the manufacture of a medicament for treatinga disease or disorder associated with a cognitive deficit.

Additionally, the invention provides a method of treating a subjectafflicted with Alzheimer's disease, comprising periodicallyadministering to the subject a pharmaceutical composition comprising anamount of pridopidine or a pharmaceutically acceptable salt thereofeffective to treat the subject.

The invention also provides pridopidine or a pharmaceutically acceptablesalt thereof for use in treating a subject afflicted with Alzheimer'sdisease.

The invention also provides pridopidine or a pharmaceutically acceptablesalt thereof for the manufacture of a medicament for use in treating asubject afflicted with Alzheimer's disease.

The invention also provides a pharmaceutical composition comprising aneffective amount of pridopidine or a pharmaceutically acceptable saltthereof for treating Alzheimer's disease.

The invention also provides a pharmaceutical composition comprisingpridopidine or a pharmaceutically acceptable salt thereof for use intreating a subject suffering from Alzheimer's disease.

The invention also provides a package comprising:

-   a) a pharmaceutical composition comprising an amount of pridopidine;    and

b) instructions for use of the pharmaceutical composition to treat asubject afflicted with a Alzheimer's disease.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with Alzheimer's disease,which comprises:

-   a) one or more unit doses, each such unit dose comprising an amount    of pridopidine thereof, wherein the amount of said pridopidine in    said unit dose is effective, upon administration to said subject, to    treat the subject, and-   b) a finished pharmaceutical container therefor, said container    containing said unit dose or unit doses, said container further    containing or comprising labeling directing the use of said package    in the treatment of said subject.

The invention also provides a package comprising:

-   -   a) a first pharmaceutical composition comprising an amount of        pridopidine and a pharmaceutically acceptable carrier;    -   b) a second pharmaceutical composition comprising an amount of a        drug approved to treat Alzheimer's disease and a        pharmaceutically acceptable carrier, and    -   c) instructions for use of the first and second pharmaceutical        compositions together to treat a subject afflicted with a        Alzheimer's disease.

The invention also provides a package comprising:

-   -   a) a first pharmaceutical composition comprising an amount of        pridopidine and a pharmaceutically acceptable carrier;    -   b) a second pharmaceutical composition comprising an amount of        one or more antidepressants, anxiolytics, or antipsychotic        medications, and a pharmaceutically acceptable carrier; and    -   c) instructions for use of the first and second pharmaceutical        compositions together to treat a subject afflicted with        Alzheimer's disease.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with Alzheimer's disease,which comprises:

-   -   a) one or more unit doses, each such unit dose comprising:        -   i) an amount of pridopidine and        -   ii) an amount of a drug approved to treat Alzheimer's            disease wherein the respective amounts of said pridopidine            and said drug approved to treat Alzheimer's disease in said            unit dose are effective, upon concomitant administration to            said subject, to treat the subject, and    -   b) a finished pharmaceutical container therefor, said container        containing said unit dose or unit doses, said container further        containing or comprising labeling directing the use of said        package in the treatment of said subject.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with Alzheimer's disease,which comprises:

-   -   a) one or more unit doses, each such unit dose comprising:        -   i) an amount of pridopidine and        -   ii) an amount of one or more antidepressants, anxiolytics,            or antipsychotic medications    -   wherein the respective amounts of said pridopidine and said one        or more antidepressants, anxiolytics, or antipsychotic        medications in said unit dose are effective, upon concomitant        administration to said subject, to treat the subject, and    -   b) a finished pharmaceutical container therefor, said container        containing said unit dose or unit doses, said container further        containing or comprising labeling directing the use of said        package in the treatment of said subject.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A and 1B: Effect of pridopidine on the Disruption of SpontaneousAlternation (FIG. 1A) and the Increase in the Number of Total ArmEntries (FIG. 1B) Induced by MK-801 in a Y-maze. Data represents themean±SEM. **p<0.01, statistically significant compared with the normalgroup (Student's t test). # p<0.05; statistically significant comparedwith the control group (Dunnett's multiple comparison test). Twelve micewere used in each group.

FIG. 2: Huntington's Disease S1R accumulates in neuronal nuclearinclusion (NNI).

FIG. 3: Pridopidine rescues spine loss in YAC128 corticostriatalco-cultures.

FIG. 4: Pridopidine rescues mushroom spine loss in PS-KI (AD) neurons.

FIG. 5: Effect of pridopidine (15 and 60 mg/kg) on Acetylcholine (Ach)levels in the PFC of rats (data expressed as mean % baseline±SEM).

FIG. 6: Effect of Pridopidine (15 and 60 mg/kg) on ACh levels in the STRof rats (data expressed as mean % baseline±SEM).

FIG. 7: Effect of pridopidine (15 and 60 mg/kg) on ACh levels in theHipp of rats (data expressed as mean % baseline±SEM).

FIG. 8: Effect of pridopidine (6 concentrations) and BDNF (50 ng/ml) onsurvival of primary neuron cultured in the presence of β amyloid 1-42(10 μM), expressed in percentage of control (mean±s.e.m; * p<0.05;**p<0.01; *** p<0.005; β amyloid 1-42 vs pridopidine/BDNF; one way Anovafollowed by Dunnett's test).

FIG. 9: Pridopidine low dose shows full SiR occupancy in a Human PETStudy and complete 18F-Fluspidine displacement after single oral dose of90 mg pridopidine.

FIG. 10: The functional connectivity within the DMN with/without 90 mgpridopidine in HV (correlates with exposure at 45 mg bid at steadystate). Upper panel: four individuals after 90 mg single dosepridopidine (post). Lower panel four individuals without pridopidinetreatment (pre).

FIG. 11: SPM-based voxelwise group comparison (within the DMN) in HVbefore (pre) and after (post) 90 mg pridopidine (correlates withexposure at 45 mg bid at steady state) reveals pridopidine increased FCin the insula.

FIG. 12: SPM-based voxelwise group comparison (within the DMN) in HDpatients before (pre) and after (post) 90 mg pridopidine (correlateswith exposure at 45 mg bid at steady state) reveals pridopidineincreased FC in the parietal and temporal cortex as well as precuneusand insula.

DETAILED DESCRIPTION OF THE INVENTION

This invention provides a method of improving cognitive function in asubject comprising periodically administering to the subject an amountof pridopidine or a pharmaceutically acceptable salt thereof effectiveto improve a cognitive function in the subject.

This invention provides a method of improving cognitive function in ahuman subject comprising administering to the subject an amount ofpridopidine or a pharmaceutically acceptable salt thereof wherein thecognitive function in need for improvement is associated with DMNdysfunction.

In an embodiment, the cognitive function is selected from the groupconsisting of global cognitive functioning, sustained cognition, memory,language, executive functioning, and attention. In another embodiment,the cognitive function is memory.

In an embodiment, memory is short term memory. In another embodiment,memory is long term memory. In another embodiment, memory is workingmemory.

In an embodiment, the subject is afflicted with a cognitive deficit. Inanother embodiment, the subject is prone to or predisposed to have acognitive deficit.

In an embodiment, the cognitive deficit is a memory deficit.

In an embodiment, the memory deficit is a short-term memory deficit. Inanother embodiment, the memory deficit is memory loss.

In an embodiment, the memory loss is caused by one or more ofage-related changes in memory, mild cognitive impairment, dementia ordepression.

In an embodiment, the cognitive deficit is caused by or associated witha disease or disorder.

In an embodiment, the disease or disorder is a disease or disorderassociated with NMDA receptor.

In another embodiment, the disease or disorder is schizophrenia orautism. In another embodiment, the disease or disorder is epilepsy or ananxiety disorder. In another embodiment, the disease or disorder isamyotrophic lateral sclerosis (ALS). In another embodiment, the diseaseor disorder is frontotemporal dementia (FTD). In another embodiment, thedisease or disorder is mild cognitive impairment (MCI). In anotherembodiment, the disease or disorder is bipolar disorder. In anotherembodiment, the disease or disorder is Huntington's disease. In anotherembodiment, the disease or disorder is selected from the groupconsisting of major depressive disorder (MDD), Parkinson's disease,Alzheimer's disease, tardive dyskinesia, depression, sickle cell anemia,stroke, chronic pain syndrome, and addiction. In another embodiment, thedisease or disorder is selected from the group consisting of mildcognitive impairment, memory loss, memory deficit, a memory deficitrelated to brain injury or a post-stroke event, a learning deficiency,and behavioral and cognitive problems associated with brain tumors. Inanother embodiment, the disease or disorder is selected from the groupconsisting of dementia, dementia associated with Lewy Bodies,age-related cognitive decline, psychosis, attention deficit disorder(ADHD), bipolar disorder, brain injury, mood and affective disorders,Tourette's syndrome, mental retardation, progressive supranuclear palsy,Creutzfeldt-Jacob disease, corticobasal Degeneration, vascular dementia,and Pick's disease. In another embodiment, the disease or disorder isselected from the group consisting of generalized anxiety disorder(GAD), social anxiety disorder (SAD), tardive dyskinesia, depression,sickle cell anemia, chronic pain syndrome, addiction, nicotineaddiction, internet addiction, cocaine addiction, tourette's syndrome,mental retardation, corticobasal degeneration, vascular dementia, pick'sdisease, posttraumatic stress disorder (PTSD), obsessive compulsivedisorder, panic disorder (PD), trigeminal pain, trigeminalmusculoskeletal pain, phantom limb pain, irritable bowel syndrome,blepharospasm, complex regional pain syndrome, chronic low back pain,autism spectrum disorder (ASD), infantile spasm (IS).

In one embodiment, this invention provides a method of treating adisease or disorder associated with DMN dysfunction comprisingadministering a pharmaceutical composition comprising pridopidine or anyacceptable salt thereof.

In an embodiment, the amount of pridopidine or a pharmaceuticallyacceptable salt thereof is administered to the human subject once daily.In one embodiment, the amount of pridopidine or a pharmaceuticallyacceptable salt thereof is administered more often than once daily. Inanother embodiment, the amount of pridopidine or a pharmaceuticallyacceptable salt thereof is administered 2-4 times a day. In oneembodiment, the amount of pridopidine or a pharmaceutically acceptablesalt thereof is administered less than once a day. In anotherembodiment, the amount of pridopidine or a pharmaceutically acceptablesalt thereof is administered 2-4 times a week.

In an embodiment, the periodic administration continues for at least 3days, more than 30 days, more than 42 days, 8 weeks or more, at least 12weeks, at least 24 weeks, more than 24 weeks, or 6 months or more.

In an embodiment, the amount of pridopidine administered is 1 mg/day-315mg/day, 22.5 mg/day-315 mg/day or 90 mg/day-315 mg/day.

In another embodiment, the amount of pridopidine administered is about22.5 mg/day, about 45 mg/day, about 67.5 mg/day, about 90 mg/day, about100 mg/day, about 112.5 mg/day, about 125 mg/day, about 135 mg/day,about 150 mg/day, about 180 mg/day, about 200 mg/day, about 250 mg/day,or about 315 mg/day.

In an embodiment, the amount of pridopidine is administered orally. Inanother embodiment, the amount of pridopidine is administeredparenterally. In another embodiment, the amount of pridopidine isadministered intravascularly. In another embodiment, the amount ofpridopidine is administered paracancerally. In another embodiment, theamount of pridopidine is administered transmucosally. In anotherembodiment, the amount of pridopidine is administered transdermally. Inanother embodiment, the amount of pridopidine is administeredintramuscularly. In another embodiment, the amount of pridopidine isadministered untranasally. In another embodiment, the amount ofpridopidine is administered intravenously. In another embodiment, theamount of pridopidine is administered intradermally. In anotherembodiment, the amount of pridopidine is administered subcutaneously. Inanother embodiment, the amount of pridopidine is administeredsublingually. In another embodiment, the amount of pridopidine isadministered intraperitoneally. In another embodiment, the amount ofpridopidine is administered intraventricularly. In another embodiment,the amount of pridopidine is administered intracranially. In anotherembodiment, the amount of pridopidine is administered intravaginally. Inanother embodiment, the amount of pridopidine is administered byinhalation. In another embodiment, the amount of pridopidine isadministered rectally. In another embodiment, the amount of pridopidineis administered intratumorally.

In an embodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine hydrochloride. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine hydrobromide. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine nitrate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine perchlorate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine phosphate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine sulphate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine formate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine acetate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine aconate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine ascorbate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine benzenesulphonate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine benzoate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine cinnamate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine citrate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine embonate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine enantate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine fumarate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidineglutamate. In another embodiment, the pharmaceutically acceptable saltof pridopidine is pridopidine glycolate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine lactate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine maleate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine malonate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine mandelate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidinemethanesulphonate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine naphthalene-2-sulphonate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine phthalate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidinesalicylate. In another embodiment, the pharmaceutically acceptable saltof pridopidine is pridopidine sorbate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine stearate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine succinate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine tartrate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine toluene-p-sulphonate.

In an embodiment, the subject is a human patient.

The invention also provides a package comprising:

a) a pharmaceutical composition comprising an amount of pridopidine or apharmaceutically acceptable salt thereof and a pharmaceuticallyacceptable carrier;

b) instructions for use of the pharmaceutical composition to treat asubject afflicted with a disease or disorder associated with a cognitivedeficit.

In an embodiment, the disease or disorder is a disease or disorderassociated with NMDA receptor.

In an embodiment, the cognitive deficit is memory loss.

In an embodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine hydrochloride.

In an embodiment, the amount of pridopidine in the pharmaceuticalcomposition is 1 mg-315 mg, 22.5 mg-315 mg or 90 mg-315 mg. In anotherembodiment, the amount of pridopidine in the pharmaceutical compositionis about 22.5 mg, about 45 mg, about 67.5, mg, about 90 mg, about 100mg, about 112.5 mg, about 125 mg, about 135 mg, about 150 mg, about 180mg, about 200 mg, about 250 mg, or about 315 mg.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with a disease or disorderassociated with a cognitive deficit, which comprises:

a) one or more unit doses, each such unit dose comprising an amount ofpridopidine or a pharmaceutically acceptable salt thereof, wherein theamount of said pridopidine in said unit dose is effective, uponadministration to said subject, to treat the subject, and

b) a finished pharmaceutical container therefor, said containercontaining said unit dose or unit doses, said container furthercontaining or comprising labeling directing the use of said package inthe treatment of said subject.

In an embodiment, the disease or disorder is a disease or disorderassociated with NMDA receptor.

The invention also provides a pharmaceutical composition comprising anamount of pridopidine or a pharmaceutically acceptable salt thereof foruse in treating a subject afflicted with a disease or disorderassociated with a cognitive deficit.

In an embodiment, the pharmaceutical composition is in a solid, acapsule or a tablet form.

In an embodiment, the cognitive deficit is memory loss.

In an embodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine hydrochloride.

In an embodiment, the amount of pridopidine in the pharmaceuticalcomposition is 1 mg-315 mg, 22.5 mg-315 mg or 90 mg-315 mg. In ananother embodiment, the amount of pridopidine in the pharmaceuticalcomposition is about 22.5 mg, about 45 mg, about 67.5, mg, about 90 mg,about 100 mg, about 112.5 mg, about 125 mg, about 135 mg, about 150 mg,about 180 mg, about 200 mg, about 250 mg, or about 315 mg.

The invention also provides a pharmaceutical composition in a unitdosage form, useful in treating a subject afflicted with a disease ordisorder associated with a cognitive deficit, which comprises an amountof pridopidine or a pharmaceutically acceptable salt thereof, whereinthe amount of said pridopidine in said composition is effective, uponadministration to said subject of one or more of said unit dosage formsof said composition, to treat the subject.

In an embodiment, the disease or disorder is a disease or disorderassociated with NMDA receptor.

The invention also provides a package comprising:

a) a pharmaceutical composition; and

b) instructions for use of the pharmaceutical composition to treat asubject afflicted with a disease or disorder associated with a cognitivedeficit.

This invention also provides pridopidine or a pharmaceuticallyacceptable salt thereof for use in treating a disease or disorderassociated with a cognitive deficit.

This invention also provides a use of pridopidine or a pharmaceuticallyacceptable salt thereof in the manufacture of a medicament for treatinga disease or disorder associated with a cognitive deficit.

For the foregoing and following embodiments, each embodiment disclosedherein is contemplated as being applicable to each of the otherdisclosed embodiments. For instance, the elements recited in the methodembodiments can be used in the pharmaceutical composition, package, anduse embodiments described herein and vice versa.

The invention further provides a method of treating a subject afflictedwith a disease selected from Alzheimer's disease and mild cognitiveimpairment (MCI), comprising periodically administering to the subject apharmaceutical composition comprising an amount of pridopidine or apharmaceutically acceptable salt thereof effective to treat the subject.

The invention further provides a method of treating a subject afflictedwith a disease selected from Alzheimer's disease and mild cognitiveimpairment (MCI), comprising periodically administering to the subject apharmaceutical composition comprising an amount of pridopidine or apharmaceutically acceptable salt thereof effective to treat the subject,wherein the disease is associated with DMN dysfunction.

In an embodiment, the amount of pridopidine is effective to reduceneurotoxicity in the subject. In another embodiment, the amount ofpridopidine is effective to inhibit the progression Alzheimer's diseaseor MCI in the subject. In a further embodiment, the amount ofpridopidine is effective to reduce one or more symptoms of Alzheimer'sdisease or MCI in the subject.

In an embodiment, the one or more symptoms are selected from the groupconsisting of cognitive impairment, function performance impairment,impairment in basic and instrumental activities of daily living, reducedquality of life and psychopathology.

In one embodiment, the one or more symptoms are measured by theClinician's Interview-based Impression of Change plus Caregiver Input(CIBIC-Plus), Severity Impairment Battery (SIB), Alzheimer's DiseaseCooperative Study Clinician's Global Impression of Change (ADCS-CCGIC),Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog), ClinicalDementia Rating (CDR), CIBIC Plus-J Score Mental Function ImpairmentScale (MENFIS), Mini-mental state exam (MMSE), Mini-cog test, BlessedInformation-Memory-Concentration Test (BIMC), CambridgeNeuropsychological Test Automated Battery (CANTAB), Alzheimer's DiseaseCooperative Study Activities of Daily Living Inventory (ADCS-ADL) Score,ADCS-ADL-SIV (severe impairment version), Disability Assessment forDementia (DAD), the Functional Assessment Questionnaire (FAQ),Instrumental Activities of Daily Living (IADL), PhysicalSelf-Maintenance Scale (PSMS) and Progressive Deterioration Scale (PDS),the Neuropsychiatric Inventory (NPI) score, CIBIC Plus-J ScoreBehavioral Pathology in Alzheimer's Disease Rating Scale (Behave-AD),CIBIC Plus-J Score Mental Function Impairment Scale (MENFIS), theResource Utilization in Dementia-Lite (RUD-Lite), EuroQol 5-DimensionalHealth-Related Quality of Life Scale (EQ-5D), the Clinical GlobalImpression of Change (CGIC), the Clinical Interview-Based Impression(CIBI), the Global Deterioration Scale (GDS), A Cognitive AssessmentBattery for Clinical Trials in Huntington's Disease (HD-CAB), MontrealCognitive Assessment (MoCA), Stroop Word Reading (SWR) or Symbol DigitModalities Test (SDMT).

In an embodiment, the periodic administration is oral.

In an embodiment, between 1 mg-315 mg, 22.5-315 mg or 90 mg-315 mgpridopidine is administered to the patient per day. In anotherembodiment, about 22.5 mg, about 45 mg, about 67.5, mg, about 90 mg,about 100 mg, about 112.5 mg, about 125 mg, about 135 mg, about 150 mg,about 180 mg, about 200 mg, about 250 mg, or about 315 mg pridopidine isadministered to the patient per day.

In an embodiment, the amount of pridopidine is administered by a unitdose of about 22.5 mg, about 45 mg, about 67.5, mg, about 90 mg, about100 mg, about 112.5 mg, about 125 mg, about 135 mg, about 150 mg, about180 mg, about 200 mg, about 250 mg, or about 315 mg pridopidine.

In an embodiment, the unit dose is administered once daily.

In an embodiment, the unit dose is administered more than once daily. Inone embodiment, the unit dose is administered 2-4 times a day. Inanother embodiment, the unit dose is administered twice per day. In oneembodiment, the unit dose is administered less than once a day. In oneembodiment, the unit dose is administered 2-4 times per week.

In an embodiment, the pridopidine is in the form of pridopidinehydrochloride.

In an embodiment, the subject is a naïve subject.

In an embodiment, the method further comprises the administration of adrug approved to treat Alzheimer's disease or MCI. In anotherembodiment, the method further comprises the administration of apsychiatric drug. In an embodiment, the drug approved to treatAlzheimer's disease is donepezil, rivastigmine, galantamine, tacrine, ormemantine.

In an embodiment, the method further comprises the administration of apsychiatric drug, an antidepressant drug, anxiolytics, antipsychoticmedications or combinations thereof.

In an embodiment, the method further comprises the administration of oneor more antidepressants. In another embodiment, the antidepressant isselected from the group consisting of citalopram, fluoxetine, paroxeine,sertraline, and trazodone.

In an embodiment, the method further comprises the administration of oneor more anxiolytics. In another embodiment, the anxiolytic is selectedfrom the group consisting of lorazepam and oxazepam.

In an embodiment, the method further comprises the administration of oneor more antipsychotic medications. In another embodiment, theantipsychotic medication is selected from the group consisting ofaripiprazole, clozapine, haloperidol, olanzapine, quetiapine,risperidone, and ziprasidone. The invention also provides pridopidine ora pharmaceutically acceptable salt thereof for use in treating a humansubject afflicted with Alzheimer's disease.

The invention provides a method of treating a human subject afflictedwith epilepsy, comprising administering to the subject a pharmaceuticalcomposition comprising an amount of pridopidine or a pharmaceuticallyacceptable salt thereof.

In an embodiment, the amount of pridopidine or a pharmaceuticallyacceptable salt thereof is administered to the human subject once daily.In one embodiment, the amount of pridopidine or a pharmaceuticallyacceptable salt thereof is administered more often than once daily.

In an embodiment, the amount of pridopidine administered is 1 mg/day-315mg/day, 22.5 mg/day-315 mg/day or 90 mg/day-315 mg/day.

In an embodiment, the amount of pridopidine is administered orally. Inanother embodiment, the amount of pridopidine is administeredparenterally. In another embodiment, the amount of pridopidine isadministered intravascularly. In another embodiment, the amount ofpridopidine is administered paracancerally. In another embodiment, theamount of pridopidine is administered transmucosally. In anotherembodiment, the amount of pridopidine is administered transdermally. Inanother embodiment, the amount of pridopidine is administeredintramuscularly. In another embodiment, the amount of pridopidine isadministered untranasally. In another embodiment, the amount ofpridopidine is administered intravenously. In another embodiment, theamount of pridopidine is administered intradermally. In anotherembodiment, the amount of pridopidine is administered subcutaneously. Inanother embodiment, the amount of pridopidine is administeredsublingually. In another embodiment, the amount of pridopidine isadministered intraperitoneally.

In another embodiment, the amount of pridopidine is administeredintraventricularly. In another embodiment, the amount of pridopidine isadministered intracranially. In another embodiment, the amount ofpridopidine is administered intravaginally. In another embodiment, theamount of pridopidine is administered by inhalation. In anotherembodiment, the amount of pridopidine is administered rectally. Inanother embodiment, the amount of pridopidine is administeredintratumorally.

In an embodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine hydrochloride.

The invention also provides pridopidine or a pharmaceutically acceptablesalt thereof for the manufacture of a medicament for use in treating asubject afflicted with Alzheimer's disease.

The invention also provides a pharmaceutical composition comprising aneffective amount of pridopidine or a pharmaceutically acceptable saltthereof for treating Alzheimer's disease.

The invention also provides a pharmaceutical composition comprisingpridopidine or a pharmaceutically acceptable salt thereof for use intreating a subject suffering from Alzheimer's disease.

The invention also provides a package comprising:

-   a) a pharmaceutical composition comprising an amount of pridopidine;    and-   b) instructions for use of the pharmaceutical composition to treat a    subject afflicted with a Alzheimer's disease.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with Alzheimer's disease,which comprises:

-   a) one or more unit doses, each such unit dose comprising an amount    of pridopidine thereof, wherein the amount of said pridopidine in    said unit dose is effective, upon administration to said subject, to    treat the subject, and-   b) a finished pharmaceutical container therefor, said container    containing said unit dose or unit doses, said container further    containing or comprising labeling directing the use of said package    in the treatment of said subject.

The method of reducing neurotoxicity in a human patient afflicted withAlzheimer's disease, comprising periodically administering to thepatient a pharmaceutical composition comprising pridopidine or apharmaceutically acceptable salt thereof effective to treat the subject.

In an embodiment, the subject is a human subject.

The invention also provides a package comprising:

-   -   a) a first pharmaceutical composition comprising an amount of        pridopidine and a pharmaceutically acceptable carrier;    -   b) a second pharmaceutical composition comprising an amount of a        drug approved to treat Alzheimer's disease and a        pharmaceutically acceptable carrier, and    -   c) instructions for use of the first and second pharmaceutical        compositions together to treat a subject afflicted with a        Alzheimer's disease.

In an embodiment, the drug approved to treat Alzheimer's disease isdonepezil, rivastigmine, galantamine, tacrine, or memantine.

The invention also provides a package comprising:

-   -   a) a first pharmaceutical composition comprising an amount of        pridopidine and a pharmaceutically acceptable carrier;    -   b) a second pharmaceutical composition comprising an amount of        one or more antidepressants, anxiolytics, or antipsychotic        medications, and a pharmaceutically acceptable carrier; and    -   c) instructions for use of the first and second pharmaceutical        compositions together to treat a subject afflicted with        Alzheimer's disease.

In an embodiment, the antidepressant is selected from the groupconsisting of citalopram, fluoxetine, paroxeine, sertraline, andtrazodone.

In an embodiment, the anxiolytic is selected from the group consistingof lorazepam and oxazepam.

In an embodiment, the antipsychotic medication is selected from thegroup consisting of aripiprazole, clozapine, haloperidol, olanzapine,quetiapine, risperidone, and ziprasidone.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with Alzheimer's disease,which comprises:

-   -   a) one or more unit doses, each such unit dose comprising:        -   i) an amount of pridopidine and        -   ii) an amount of a drug approved to treat Alzheimer's            disease        -   wherein the respective amounts of said pridopidine and said            drug approved to treat Alzheimer's disease in said unit dose            are effective, upon concomitant administration to said            subject, to treat the subject, and    -   b) a finished pharmaceutical container therefor, said container        containing said unit dose or unit doses, said container further        containing or comprising labeling directing the use of said        package in the treatment of said subject.

In an embodiment, the drug approved to treat Alzheimer's disease isdonepezil, rivastigmine, galantamine, tacrine, or memantine.

The invention also provides a therapeutic package for dispensing to, orfor use in dispensing to, a subject afflicted with Alzheimer's disease,which comprises:

-   -   a) one or more unit doses, each such unit dose comprising:        -   i) an amount of pridopidine and        -   ii) an amount of one or more antidepressants, anxiolytics,            or antipsychotic medications        -   wherein the respective amounts of said pridopidine and said            one or more antidepressants, anxiolytics, or antipsychotic            medications in said unit dose are effective, upon            concomitant administration to said subject, to treat the            subject, and    -   b) a finished pharmaceutical container therefor, said container        containing said unit dose or unit doses, said container further        containing or comprising labeling directing the use of said        package in the treatment of said subject.

In an embodiment, the antidepressant is selected from the groupconsisting of citalopram, fluoxetine, paroxeine, sertraline, andtrazodone.

In an embodiment, the anxiolytic is selected from the group consistingof lorazepam and oxazepam.

In an embodiment, the antipsychotic medication is selected from thegroup consisting of aripiprazole, clozapine, haloperidol, olanzapine,quetiapine, risperidone, and ziprasidone.

Combinations of the above-described embodiments are also within thescope of the invention.

The antipsychotic medication may be used to treat hallucinations,delusions, aggression, agitation, hostility and/or uncooperativeness.The anxiolytic may be used to treat anxiety, restlessness, verballydisruptive behavior and/or resistance. The antidepressant may be used totreat low mood and irritability (www.alz.org).

For the foregoing embodiments, each embodiment disclosed herein iscontemplated as being applicable to each of the other disclosedembodiments. In addition, the elements recited in pharmaceuticalcomposition embodiments can be used in the method and use embodimentsdescribed herein.

Terms

As used herein, and unless stated otherwise, each of the following termsshall have the definition set forth below.

As used herein, “administering to the subject” means the giving of,dispensing of, or application of medicines, drugs, or remedies to asubject to relieve, cure or reduce the symptoms associated with adisease, disorder or condition, e.g., a pathological condition. Oraladministration is one way of administering the instant compounds to thesubject.

As used herein, “cognitive function” means an intellectual process bywhich one becomes aware of, perceives, or comprehends ideas. Cognitivefunction involves all aspects of perception, thinking, reasoning, andmemory, including short term memory.

As used herein, “improving cognitive function” includes slowing,stopping, or reversing the progression of a cognitive deficit, inaddition to increasing cognitive function. Areas of cognitive functionare described in Fioravanti et al (2012).

As used herein, a “cognitive deficit” is an inclusive term to describeany characteristic that acts as a barrier to cognitive function.Cognitive deficits can include loss of higher reasoning, forgetfulness,learning disabilities, concentration difficulties, decreasedintelligence, and other reductions in mental functions. Cognitivedeficits may be congenital or caused by environmental factors, braininjuries, neurological disorders, or mental illness. Cognitiveimpairments and cognitive dysfunctions are also considered cognitivedeficits.

As used herein, “short term memory” is the capacity to recognize, recalland regurgitate small amounts of information shortly after itsoccurrence.

MK-801, or Dizocilpine, is an NMDA receptor antagonist.

As used herein, a “disease or disorder associated with an NMDA receptor”is any disease or disorder related to or resulting from NMDA receptorimbalance or dysfunction. This can include, but is not limited to,diseases or disease symptoms that can be induced or caused by NMDAreceptor antagonists such as MK-801, diseases which may be treated byNMDA receptor antagonists such as MK-801, or diseases in which NMDAreceptor antagonists such as MK-801 increase or decrease the severity ofthe symptoms.

The methods of the present invention are useful for improving cognitivefunction in diseases and disorders associated with both cognitivedeficits and the NMDA receptor. The following diseases and disorders areassociated with cognitive deficits and the NMDA receptor: schizophrenia,autism (Saunders, 2012), epilepsy (van Rijckevorsel 2006, Stafstrom1997), anxiety disorders (Ferreri 2011, Dietz 2014), major depressivedisorder (MDD) (Keefe 2014, Rapaport 2009), Parkinson's disease (Dubois1997, Jonkers 2000), Alzheimer's disease (U.S. Patent Publication No.20130065966), tardive dyskinesia (Krabbendam 2000, Seo 1997), Depression(Austin 2001, Ates-Alagoz 2013), sickle cell anemia (Steen 2005, U.S.Pat. No. 8,680,042), stroke (Cumming 2013, U.S. Patent Publication No.20130065966), chronic pain syndrome (Hart 2003, Zeynep 2013), addiction(Gould 2010, U.S. Pat. No. 5,321,012), and Huntington's disease (Foroud1995, U.S. Patent Publication No. 20130065966).

Other diseases and disorders which may be treated by the methods of thisinvention include: memory deficit, mild cognitive impairment, memoryloss, a memory deficit related to brain injury or a post-stroke event, alearning deficiency, and behavioral and cognitive problems associatedwith brain tumors.

Additional diseases and disorders which may be treated by the methods ofthis invention include: dementia, dementia associated with Lewy Bodies,age-related cognitive decline, psychosis, attention deficit disorder(ADHD), bipolar disorder, brain injury, mood and affective disorders,Tourette's syndrome, mental retardation, progressive supranuclear palsy,Creutzfeldt-Jacob disease, vascular dementia, Corticobasal Degeneration,Creutzfeldt-Jakob Disease, and Pick's disease.

As used herein, an “amount” or “dose” of pridopidine as measured inmilligrams refers to the milligrams of pridopidine(4-[3-(methylsulfonyl)phenyl]-1-propyl-piperidine) present in apreparation, regardless of the form of the preparation. For example, aunit dose containing “90 mg pridopidine” means the amount of pridopidinebase in a preparation is 90 mg, regardless of the form of thepreparation. Thus, when in the form of a salt, e.g. pridopidinehydrochloride salt, the weight of the salt form necessary to provide adose of 90 mg pridopidine would be greater than 90 mg due to thepresence of the salt.

As used herein, a “unit dose”, “unit doses” and “unit dosage form(s)”mean a single drug administration entity/entities.

As used herein, “about” in the context of a numerical value or rangemeans±10%/o of the numerical value or range recited or claimed.

As used herein, “effective” as in an amount effective to achieve an endmeans the quantity of a component that is sufficient to yield anindicated therapeutic response without undue adverse side effects (suchas toxicity, irritation, or allergic response) commensurate with areasonable benefit/risk ratio when used in the manner of thisdisclosure. For example, an amount effective to treat cognitive deficit.The specific effective amount varies with such factors as the particularcondition being treated, the physical condition of the patient, the typeof mammal being treated, the duration of the treatment, the nature ofconcurrent therapy (if any), and the specific formulations employed andthe structure of the compounds or its derivatives.

As used herein, to “treat” or “treating” encompasses, e.g., inducinginhibition, regression, or stasis of a disorder and/or disease, e.g. AD,or alleviating, lessening, suppressing, inhibiting, reducing theseverity of, eliminating or substantially eliminating, or ameliorating asymptom of the disease or disorder.

As used herein, “inhibition” of disease progression or diseasecomplication in a subject means preventing, delaying or reducing thedisease progression and/or disease complication in the subject.

This includes, for example, delaying the progression of one of moresymptoms in the subject, including for example delaying the progressionof cognitive impairment, delaying the deterioration in functionperformance and basic and instrumental activities of daily living,delaying the deterioration of quality of life or delaying thedeterioration of psychopathology.

A “symptom” associated with AD includes any clinical or laboratorymanifestation associated with AD and is not limited to what the subjectcan feel or observe. Symptoms of AD include but are not limited to theimpairment and/or deterioration in cognition (e.g., memory and behavior,judgment/problem solving, attention, concentration, naming,comprehension, reasoning, language, communication, orientation andpraxis), functional performance (e.g., grooming, dressing, walkingincluding balance, bathing, feeding and toileting) basic andinstrumental activities of daily living (e.g., shopping, preparingmeals, using household appliances, conducting hobbies and interests,keeping appointments and reading), quality of life (e.g., mobility,self-care, daily activities, pain/discomfort, mood, relationships,overall physical condition, anxiety and depression, swallowing), orpsychopathology (e.g. paranoid and delusional ideation, hallucinations,activity disturbances, diurnal rhythm disturbances, aggressiveness,affective disorders and anxieties, and phobias).

Various assessment tools are accepted in the field, which serve toevaluate the status of AD patients. For example, Cognition may beevaluated by various assessment tools exemplified by the ClinicianInterview-Based Impression of Change, plus career interview(CIBIC-Plus), Severity Impairment Battery (SIB), Alzheimer's DiseaseCooperative Study Clinician's Global Impression of Change (ADCS-CCGIC),Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog), ClinicalDementia Rating (CDR), CIBIC Plus-J Score Mental Function ImpairmentScale (MENFIS), Mini-mental state exam (MMSE), Mini-cog test, BlessedInformation-Memory-Concentration Test (BIMC), and CambridgeNeuropsychological Test Automated Battery (CANTAB). Functionalperformance and basic and instrumental activities of daily living can beevaluated for example using Alzheimer's Disease Cooperative StudyActivities of Daily Living Inventory (ADCS-ADL) Score, ADCS-ADL-SIV(severe impairment version), Disability Assessment for Dementia (DAD),Functional Assessment Questionnaire (FAQ), Instrumental Activities ofDaily Living (IADL), Physical Self-Maintenance Scale (PSMS) andProgressive Deterioration Scale (PDS). Psychopathology can be evaluatedfor example by Neuropsychiatric Inventory (NPI) score, CIBIC Plus-JScore Behavioral Pathology in Alzheimer's Disease Rating Scale(Behave-AD), CIBIC Plus-J Score Mental Function Impairment Scale(MENFIS). Quality of life can be evaluated for example by the ResourceUtilization in Dementia-Lite (RUD-Lite), EuroQol 5-DimensionalHealth-Related Quality of Life Scale (EQ-SD), including for exampleProxy Version (EQ-SD Proxy). In addition, global assessment measures maybe used to evaluate the patient's status, indulging for example ClinicalGlobal Impression of Change (CGIC), Clinical Interview-Based Impression(CIBI), Global Deterioration Scale (GDS).

A “biomarker” is a measurable parameter that serves as an indication tothe severity, subtype or stage of the disease. Biomarkers for AD includebut are not limited to Amyloid Beta (Aβ) 1-42 plasma concentration,change in glucose metabolism in the brain measured for example byPositron Emission Tomography (PET), and change in hyppocampal volume,measured for example by Magnetic Resonance Imaging (MRI). An increasedAmyloid Beta (Aβ) 1-42 plasma concentration, reduced glucose metabolismin the brain, and reduced hyppocampal volume are considered to beassociated with increased severity of the disease.

As used herein, “a subject afflicted with AD” means a subject diagnosedas suffering from AD, including for example subjects diagnosed assuffering from “definite AD”, “probable AD” or “possible AD”. In anembodiment, the subject is diagnosed as according to the 1984 Criteria,also called the NINCDS-ADRDA Alzheimer's criteria (McKhann et al. 1984).In another embodiment, the subject is diagnosed according to the revisedcriteria for diagnosis of Alzheimer's disease of the National Instituteon Aging-Alzheimer's Association diagnostic guidelines for Alzheimer'sdisease (McKhann et al. 2011). In another embodiment, the subject isdiagnosed according to the DSM-IV criteria. In another embodiment, thesubject is diagnosed according to the International Classification ofDiseases.

“Adverse event” or “AE” means any untoward medical occurrence in aclinical trial subject administered a medicinal product and which doesnot have a causal relationship with the treatment. An adverse event cantherefore be any unfavorable and unintended sign including an abnormallaboratory finding, symptom, or diseases temporally associated with theuse of an investigational medicinal product, whether or not consideredrelated to the investigational medicinal product.

As used herein, a subject at “baseline” is a subject prior to initiatingperiodic administration of pridopidine.

As used herein, a “naïve subject” or a “naïve patient” with respect to adrug or therapy means that the subject has not previously received thedrug or therapy.

A “pharmaceutically acceptable carrier” refers to a carrier or excipientthat is suitable for use with humans and/or animals without undueadverse side effects (such as toxicity, irritation, and allergicresponse) commensurate with a reasonable benefit/risk ratio. It can be apharmaceutically acceptable solvent, suspending agent or vehicle, fordelivering the instant compounds to the subject.

As used herein, the term “pridopidine” refers to pridopidine free base.In certain embodiments, pridopidine also includes any pharmaceuticallyacceptable salt, such as the HCl salt or tartrate salt. Preferably, inany embodiments of the invention as described herein, the pridopidine isin the form of its hydrochloride salt.

As used herein, “combination” means an assemblage of reagents for use intherapy either by simultaneous or contemporaneous administration.Simultaneous administration refers to administration of an admixture(whether a true mixture, a suspension, an emulsion or other physicalcombination) of the pridopidine and the second compound. In this case,the combination may be the admixture or separate containers of thepridopidine and the second compound that are combined just prior toadministration. Contemporaneous administration refers to the separateadministration of the pridopidine and the second compound at the sametime, or at times sufficiently close together that an additive orpreferably synergistic activity relative to the activity of either thethe pridopidine and the second compound alone is observed.

As used herein, “concomitant administration” or administering“concomitantly” means the administration of two agents given in closeenough temporal proximately to allow the individual therapeutic effectsof each agent to overlap.

As used herein, “add-on” or “add-on therapy” means an assemblage ofreagents for use in therapy, wherein the subject receiving the therapybegins a first treatment regimen of one or more reagents prior tobeginning a second treatment regimen of one or more different reagentsin addition to the first treatment regimen, so that not all of thereagents used in the therapy are started at the same time. For example,adding pridopidine therapy to a patient already receiving donepeziltherapy.

A dosage unit as used herein may comprise a single compound or mixturesof compounds thereof. A dosage unit can be prepared for oral dosageforms, such as tablets, capsules, pills, powders, and granules.

Pharmaceutically Acceptable Salts

The active compounds for use according to the invention may be providedin any form suitable for the intended administration. Suitable formsinclude pharmaceutically (i.e. physiologically) acceptable salts, andpre- or prodrug forms of the compound of the invention.

In an embodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine hydrochloride. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine hydrobromide. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine nitrate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine perchlorate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine phosphate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine sulphate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine formate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine acetate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine aconate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine ascorbate. In anotherembodiment, the pharmaceutically acceptable salt of pridopidine ispridopidine benzenesulphonate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine benzoate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine cinnamate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine citrate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine embonate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine enantate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine fumarate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidineglutamate. In another embodiment, the pharmaceutically acceptable saltof pridopidine is pridopidine glycolate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine lactate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine maleate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine malonate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine mandelate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidinemethanesulphonate. In another embodiment, the pharmaceuticallyacceptable salt of pridopidine is pridopidine naphthalene-2-sulphonate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine phthalate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidinesalicylate. In another embodiment, the pharmaceutically acceptable saltof pridopidine is pridopidine sorbate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine stearate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine succinate. In another embodiment, thepharmaceutically acceptable salt of pridopidine is pridopidine tartrate.In another embodiment, the pharmaceutically acceptable salt ofpridopidine is pridopidine toluene-p-sulphonate.

Such salts may be formed by procedures well known and described in theart.

Pharmaceutical Compositions

While the compounds for use according to the invention may beadministered in the form of the raw compound, it is preferred tointroduce the active ingredients, optionally in the form ofphysiologically acceptable salts, in a pharmaceutical compositiontogether with one or more adjuvants, excipients, carriers, buffers,diluents, and/or other customary pharmaceutical auxiliaries.

In an embodiment, the invention provides pharmaceutical compositionscomprising the active compounds or pharmaceutically acceptable salts orderivatives thereof, together with one or more pharmaceuticallyacceptable carriers therefore, and, optionally, other therapeutic and/orprophylactic ingredients know and used in the art. The carrier(s) mustbe “acceptable” in the sense of being compatible with the otheringredients of the formulation and not harmful to the recipient thereof.

The pharmaceutical composition of the invention may be administered byany convenient route, which suits the desired therapy. Preferred routesof administration include oral administration, in particular in tablet,in capsule, in dragé, in powder, or in liquid form, and parenteraladministration, in particular cutaneous, subcutaneous, intramuscular, orintravenous injection. The pharmaceutical composition of the inventioncan be manufactured by the skilled person by use of standard methods andconventional techniques appropriate to the desired formulation. Whendesired, compositions adapted to give sustained release of the activeingredient may be employed.

Further details on techniques for formulation and administration may befound in the latest edition of Remington's Pharmaceutical Sciences(Maack Publishing Co., Easton, Pa.).

It is understood that where a parameter range is provided, all integerswithin that range, and tenths thereof, are also provided by theinvention. For example, “0.1 mg-40.0 mg” includes 0.1 mg, 0.2 mg, 0.3mg, 0.4 mg, etc. up to 40.0 mg.

This invention will be better understood by reference to theExperimental Details which follow, but those skilled in the art willreadily appreciate that the specific experiments detailed are onlyillustrative of the invention as described more fully in the claimswhich follow thereafter.

EXPERIMENTAL DETAILS Example 1: Pridopidine Attenuates theMK-801-Induced Deficit of Working Memory in Y-Maze Mice

This Example examines the effect of pridopidine on the deficits ofworking memory and the increase in total arm entries induced by MK-801in the mouse Y-maze.

Materials and Methods

Animals: Male ddY mice (Japan SLC Inc., Shizuoka, Japan) aged 5 weekswere used. The mice were maintained with free access to laboratory chowand water. The mice were placed in the experiment room at least 1 hbefore the experiment.

Drugs: Pridopidine hydrochloride was dissolved in water and administeredorally. (+)-MK-801 hydrogen maleate (dizocilpine hydrogen maleate)(commercially available from Sigma-Aldrich (St. Louis, Mo., USA)) wasdissolved in saline and administered intraperitoneally. The drugs wereadministered at a volume of 10 mL/kg. The dose levels of pridopidine arepresented as the amount of free base.

Y-maze: The Y-maze is a learning and memory test for rodents (van denBuuse 2005). Y-maze test is based on the innate curiosity of rodents toexplore novel environments (Luszczki et al., 2005). It is used to assessexploratory behaviours, learning and memory function, short term memory,working memory, general locomotor activity, and stereotypic behavior inrodents (Hazim 2011, Onaolapo 2012, Detrait 2010).

Rodents exhibit a tendency to alternate between maze arms, therebyproviding a measure of short term spatial memory. A high alternationrate is indicative of sustained cognition as the animals must rememberwhich arm was entered last to not reenter it (Bryan 2009).

The Y-maze was made of gray vinyl chloride. Each arm was 40 cm long, 13cm high, 3 cm wide at the bottom, and 10 cm wide at the top. All armsconverged at equal angles.

Experimental Procedure: The present experiments were carried outaccording to the method established by Maurice et al. (1997).Pridopidine was administered, followed by MK-801 10 minutes later.Twenty minutes after administration of MK-801, each mouse was placed atthe end of one arm and allowed to move freely through the maze during an8-minute session. The series of arm entries were recorded. Alternationwas defined as visits into all three arms on consecutive occasions. Thenumber of maximum alternations was the total number of arm entries minus2. The percentage of alternation was calculated as the number ofalternations divided by maximum alternations and multiplied by 100.

Statistical Analysis: The percentage of alternation and total armentries were expressed as the mean±SEM. The statistical significancebetween the normal group and the control group was calculated using theStudent's t test. The statistical analysis for drug treatment versus thecontrol group was conducted using Dunnett's multiple comparison test.The level of statistical significance was p<0.05.

Results

Administration of MK-801 significantly (P<0.01, FIG. 1A, using theStudent's t test) decreased the alternation percentage. Pridopidinerestored the alternation percentage in the MK-801-treated mice withstatistical significance at 10 and 20 mg/kg (P<0.05, FIG. 1A, byDunnett's multiple comparison test). The total number of arm entries wassignificantly (P<0.01, FIG. 1B, using Student's t test) augmented bytreatment with MK-801. Pridopidine showed no statistically significantchange in the MK-801-induced increase in the total number of arm entries(FIG. 1B).

Discussion

In the Y-maze, the non-competitive NMDA antagonist, MK-801(dizocilpine), impairs spontaneous alternation behavior, which reflectsworking memory, and enhances the total number of arm entries, whichrepresents locomotor activity (Parada-Turska and Turski, 1990).

Example 1 demonstrates the anti-amnesic effects of pridopidine onworking memory impairment related to the NMDA receptor blockade.Pridopidine attenuated the MK-801-induced decrease in percentalternation with statistical significance at 10 and 20 mg/kg. Theseresults demonstrate that pridopidine improves cognitive status andimproves cognitive impairment (such as working memory) in patients,including patients with cognitive deficits.

Nilsson 2004 showed that pridopidine counteracted the behavioralprimitivization induced by MK-801 in mice. However, the experimentscompleted in Nilsson 2004 are different from the experiment described inExample 1. Specifically, the experiments completed in Nilsson 2004 didnot show that pridopidine can improve cognitive function or memory loss.

Example 1 uses a Y-maze which, as described above, is a commonly usedexperiment to measure cognitive and memory function. In the Y-mazeexperiment, the ability of the rodent to use its cognitive function toremember which arm of the Y-maze it traveled to previously is testedwhen measuring percentage of alternation. In comparison, the experimentin Nilsson 2004 (and further described in Nilsson 2001) does not containsuch a memory component. The experiment in Nilsson 2004 placed rodentsin a rectangle shaped arena and used a camera to measure the movementsand behavior of the mice. The specific behaviors measured were forwardlocomotion, rearing with exploratory sniffing, grooming and digging. InNilsson 2004 and Nilsson 2001 treating mice with MK-801 prior to theexperiment causes mice that naturally alternate between the specificmeasured behaviors to mostly perform monotonous forward locomotion(Nilsson 2001). What Nilsson 2004 found was that pridopidinecounteracted such effects of MK-801 on mice. However, there is no clearrelationship between the measured behaviors and cognitive function inthe experiment of Nilsson 2004. Thus, the experiments shown in Nilsson2004 cannot show that pridopidine enhances cognitive function. This islikely why the authors of Nilsson 2004 state that additional tests arenecessary to determine if pridopidine can enhance cognitive function.

Example 2: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Schizophrenia

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with schizophrenia.Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective to reduce cognitive deficits associated with schizophreniain the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from schizophrenia. The administration of thecomposition is effective to treat the subject suffering fromschizophrenia. The administration of the composition is also effectiveto reduce cognitive deficits associated with schizophrenia in thesubject.

Example 3: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Autism

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with autism. Periodic(e.g., daily or twice daily) oral administration of pridopidine iseffective to reduce cognitive deficits associated with autism in thesubject.

A pridopidine composition as described herein is administered orally toa subject suffering from autism. The administration of the compositionis effective to treat the subject suffering from autism. Theadministration of the composition is also effective to reduce cognitivedeficits associated with autism in the subject.

Example 4: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Epilepsy

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with epilepsy.Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective to reduce cognitive deficits associated with epilepsy inthe subject.

A pridopidine composition as described herein is administered orally toa subject suffering from epilepsy. The administration of the compositionis effective to treat the subject suffering from epilepsy. Theadministration of the composition is also effective to reduce cognitivedeficits associated with epilepsy in the subject.

Example 5: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Anxiety Disorders

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with anxietydisorders. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withanxiety disorders in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from an anxiety disorder. The administration of thecomposition is effective to treat the subject suffering from an anxietydisorder. The administration of the composition is also effective toreduce cognitive deficits associated with an anxiety disorder in thesubject.

Example 6: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Major Depressive Disorder

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with major depressivedisorder. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withmajor depressive disorder in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from major depressive disorder. The administrationof the composition is effective to treat the subject suffering frommajor depressive disorder. The administration of the composition is alsoeffective to reduce cognitive deficits associated with an anxietydisorder in the subject.

Example 7: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Parkinson's Disease

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with Parkinson'sdisease. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withParkinson's disease in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from Parkinson's disease. The administration of thecomposition is effective to treat the subject suffering from Parkinson'sdisease. The administration of the composition is also effective toreduce cognitive deficits associated with Parkinson's disease in thesubject.

Example 8: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Alzheimer's Disease

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with Alzheimer'sdisease. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withAlzheimer's disease in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from Alzheimer's disease. The administration of thecomposition is effective to treat the subject suffering from Alzheimer'sdisease. The administration of the composition is also effective toreduce cognitive deficits associated with Alzheimer's disease in thesubject.

Example 9: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Tardive Dyskinesia

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with tardivedyskinesia. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withtardive dyskinesia in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from tardive dyskinesia. The administration of thecomposition is effective to treat the subject suffering from tardivedyskinesia. The administration of the composition is also effective toreduce cognitive deficits associated with tardive dyskinesia in thesubject.

Example 10: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Depression

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with Depression.Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective to reduce cognitive deficits associated with Depression inthe subject.

A pridopidine composition as described herein is administered orally toa subject suffering from Depression. The administration of thecomposition is effective to treat the subject suffering from Depression.The administration of the composition is also effective to reducecognitive deficits associated with Depression in the subject.

Example 11: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Sickle Cell Anemia

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with sickle cellanemia. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withsickle cell anemia in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from sickle cell anemia. The administration of thecomposition is effective to treat the subject suffering from sickle cellanemia. The administration of the composition is also effective toreduce cognitive deficits associated with sickle cell anemia in thesubject.

Example 12: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Stroke

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients after having a stroke. Periodic(e.g., daily or twice daily) oral administration of pridopidine iseffective to reduce cognitive deficits associated with stroke in thesubject.

A pridopidine composition as described herein is administered orally toa subject after suffering from a stroke. The administration of thecomposition is effective to treat the subject after suffering from astroke. The administration of the composition is also effective toreduce cognitive deficits associated with stroke in the subject.

Example 13: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Chronic Pain Syndrome

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with chronic painsyndrome. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withchronic pain syndrome in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from chronic pain syndrome. The administration ofthe composition is effective to treat the subject suffering from chronicpain syndrome. The administration of the composition is also effectiveto reduce cognitive deficits associated with chronic pain syndrome inthe subject.

Example 14: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Addiction

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with addiction.Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective to reduce cognitive deficits associated with addiction inthe subject.

A pridopidine composition as described herein is administered orally toa subject suffering from addiction. The administration of thecomposition is effective to treat the subject suffering from addiction.The administration of the composition is also effective to reducecognitive deficits associated with addiction in the subject.

Example 15: Assessment of Efficacy of Pridopidine for Treating PatientsAfflicted with Huntington's Disease

Periodic (e.g., daily or twice daily) oral administration of pridopidineis effective in treating human patients afflicted with Huntington'sdisease. Periodic (e.g., daily or twice daily) oral administration ofpridopidine is effective to reduce cognitive deficits associated withHuntington's disease in the subject.

A pridopidine composition as described herein is administered orally toa subject suffering from Huntington's disease. The administration of thecomposition is effective to treat the subject suffering fromHuntington's disease. The administration of the composition is alsoeffective to reduce cognitive deficits associated with Huntington'sdisease in the subject.

Example 16

Alteration in the density of spines and abnormalities in the size andshape of spines was observed in the brains of HD patients (Graveland1985).

FIG. 3 shows that pridopidine rescues spine loss in YAC128corticostriatal co-cultures. FIG. 4 shows that pridopidine rescuesmushroom spine loss in PS-KI (AD) neurons.

Dendritic spines are important in maintaining cognition and motorfunctions. Dendritic spine formation is critical for the establishmentof excitatory synaptic networks. Spines show structural plasticity asthe basis for the physiological changes in synaptic efficacy thatunderlie learning and memory. Motor control is regulated bycortical-medium spiny neurons synaptic connections. The spines are thebasis for these connections (Kreitzer 2008 and Bourne 2008).

Example 17: Acetylcholine and Pridopidine

Animals: Eighteen adult male Sprague Dawley rats (Harlan, USA) wereused. Before the experiment, rats were group housed in plastic cages(3-4 animals/cage) and had access to food and water ad libitum. Animalswere kept on a 12/12 hour light/dark cycle. Experiments were conductedin accordance with the protocols approved by the Institutional AnimalCare and Use Committee of Brains On-Line, LLC.

Microdialysis Experiment: Microdialyis experiments were performed oneday after surgery. On the day of the experiment, the probes wereconnected with flexible PEEK tubing to a microperfusion pump (HarvardPHD 2000 Syringe pump, Holliston, Mass. or similar). Microdialysisprobes were perfused with aCSF containing 147 mM NaCl, 3.0 mM KCl, 1.2mM CaCl2) and 1.2 mM MgCl2, at a flow rate of 1.5 μL/min. Microdialysissamples were collected in 30 minute periods by an automated fractioncollector (820 Microsampler, Univentor, Malta) into 300 μL polystryenemini-vials already containing 15 μL 0.02 M formic acid (FA) and 0.04%ascorbic acid in ultrapurified H₂O. Four basal samples were collectedbefore pridopidine (15 or 60 mg/kg, PO) or vehicle was administered.Samples were collected.

Results: Acetylcholine (Ach) Levels in Dialysate

FIGS. 5, 6, and 7 show the levels of ACh in the prefrontal cortex (PFC),striatum (STR), and hippocampus (Hipp), respectively. Treatment withpridopidine resulted in significant increases in Ach in the prefrontalcortex following 15 mg/kg administration at 30 min after administration(219% change from baseline; p<0.05), at 60 min after administration(226% change from baseline; p<0.05) and at 90 min after administration(226% change from baseline; p<0.05), and following 60 mg/kgadministration at 30 min after administration (215% change frombaseline; p<0.05), at 60 min after administration (201% change frombaseline; p<0.05), at 90 min after administration (211% change frombaseline; p<0.05) and at 180 min after administration (212% change frombaseline; p<0.05). Treatment with pridopidine resulted in significantincreases in Ach in the hippocampus following 15 mg/kg administration at30 min after administration (160% change from baseline; p<0.05) and at60 min after administration (173% change from baseline; p<0.05), andfollowing 60 mg/kg administration at 60 min after administration (157%change from baseline; p<0.05) and at 90 min after administration (158%change from baseline; p<0.05) Tables 1 and 2 present the Maximum %Change from baseline in Ach levels after 15 and 60 mg/kg administration,respectively.

TABLE 1 Acetylcholine levels in dialysate (% Change from baseline.Single dose 15 mg/kg p.o) Prefrontal Striatum cortex HippocampusAcetylcholine 107 226* 173* *statistically significant (p < 0.05)

TABLE 2 Acetylcholine levels in dialysate (% Change from baseline.Single dose 60 mg/kg p.o) Prefrontal Striatum cortex HippocampusAcetylcholine 120 215* 158* *statistically significant (p < 0.05)

Acetylcholine is an important neurotransmitter in the nervous system andis necessary for learning and memory function (Winkler 1995).Acetylcholine has been accorded an important role in supporting learningand memory processes in the hippocampus. Cholinergic activity in thehippocampus and prefrontal cortex is correlated with memory (Hironaka2001). Additionally, restoration of ACh in the hippocampus afterdisruption of the septohippocampal pathway is sufficient to rescuememory (Parent 2004).

Example 18: Effect of Pridopidine in a Model of AD: Injury of RatPrimary Cortical Neurons by Human Amyloid Beta 1-42

AD is characterized by the progressive accumulation of intracellularneurofibrillary tangles, extracellular parenchymal senile plaques, andcerebrovascular deposits comprised of amyloid-β 1-42 peptides (Sakono etal, 2010).

A β peptide is a proteolytic product derived through sequentialproteolysis of amyloid precursor protein (APP), which occurs as a resultof cleavage by β-secretase and γ-secretase. Mutations at the cleavagesites in APP increase the production of A β oligomers. The progressiveaccumulation of Aβ in the form of senile plaques, which is one of thepathologic hallmarks of AD, had been recognized as one of the majorcauses of AD pathology (Kawahara and Kuroda, 2000) by triggeringneurotoxicity, oxidative damage, and inflammation (Pike et al., 1991;Cummings et al., 1998; Combs et al., 2000). The most abundant Aβ peptideform found in AD brain senile plaques are the 40 and 42 amino acid forms(Sisodia et al., 1990).

However, the number of senile plaques in a particular region of the ADbrain correlates poorly with the local extent of neuron death orsynaptic loss, or with cognitive impairment. Recent studies show arobust correlation between the soluble Aβ oligomer (AβO) levels and theextent of synaptic loss and severity of cognitive impairment (for reviewsee Sakono et al., 2010).

The study investigated the neuroprotective effect of pridopidine oncortical neurons incubated for 24 hours in the presence of AβO, an invitro model of AD (Callizot et al., 2013). BDNF at 50 ng/ml was used asa positive control in this study.

Experimental Protocol

Cortical Neurons Cell Culture

Rat cortical neurons were cultured as follows. Pregnant female rats of15 days gestation were killed by cervical dislocation (Rats Wistar;Janvier Lab) and the fetuses were removed from the uterus. The cortexeswere removed and placed in ice-cold medium of Leibovitz (L15,Panbiotech, ref: P04-27055) containing 2% of Penicillin 10.000 U/ml andStreptomycin 10 mg/ml (PS, Panbiotech, ref: P06-07100) and 1% of BovineSerum Albumin (BSA, Panbiotech, Ref: P06-1391100). Cortexes weredissociated by trypsin-EDTA (Panbiotech, Ref: P10-023100) for 20 min at37° C. The reaction was stopped by the addition of Dulbecco's modifiedEagle's Medium (DMEM, Panbiotech, Ref P04-03600) containing DNasel gradeII (0.1 mg/ml, Panbiotech, ref: P60-37780100) and 10% of Foetal CalfSerum (FCS, Invitrogen, ref: 10270-098). Cells were then mechanicallydissociated by 3 serial passages through a 10 ml pipette. Cells werethen centrifuged at 515×g for 10 min at 4° C. The supernatant wasdiscarded and the pellet of cells was re-suspended in a defined culturemedium consisting of Neurobasal (Nb, Invitrogen, ref: 21103)supplemented with B27 (20%, Invitrogen, ref: 17504), L-glutamine (2 mM,Panbiotech, ref: P04-80100), 2% of PS solution and 10 ng/ml of ofBrain-derived neurotrophic factor (BDNF, PanBiotech, Ref: CB-1115002).Viable cells were counted in a Neubauer cytometer using the trypan blueexclusion test. The cells were seeded at a density of 30,000 cells/wellin 96 well-plates pre-coated with poly-L-lysine (Greiner ref: 655930)and were cultured at +37° C. in a humidified air (95%)/CO₂ (5%)atmosphere.

After 11 days of culture, cortical neurons were intoxicated with humanAβ 1-42 at 10 μM for 24 hrs.

One culture was performed per condition, 6 wells per condition.

Pridopidine and Human β Amyloid 1-42 Treatment

The human β amyloid 1-42 peptide preparation was done following aninternal and original procedure validated by Neuron experts (Callizot etal., 2013). Briefly, on day 12 of culture, the supernatant was removedand fresh medium was added, with human amyloid β 1-42 (10 μM) and withpridopidine.

The following conditions were included:

a) Medium without Human β amyloid 1-42 (control) for 24 h

b) Human β amyloid 1-42 (10 μM) for 24 h

c) Human β amyloid 1-42 (10 μM) for 24 h with pridopidine (200, 50, 10,1, 0.1 and 0.01 μM)

d) Human β amyloid 1-42 (10 μM) for 24 h with BDNF (50 ng/ml)

End Point Evaluation: Measure of Total Number of MAP 2 Neurons.

Neurons that survived after 24 h incubation were stained with MAP 2.

After 24 hours, cells were washed twice in Phosphate Buffered Saline(PBS, PanBiotech, ref: P04-36500) and then fixed by a cold solution ofethanol (95%, Sigma, ref: 32221) and acetic acid (5%, Sigma, ref: 33209)for 5 min. The cells were then permeabilized and non-specific sites wereblocked with a solution of PBS containing 0.1% of saponin (SigmaAldrich, ref: S7900) and 1% FCS for 15 min at room temperature. Then,cells were incubated for 2 hours with mouse monoclonal primaryMicrotubule-associated protein 2 antibody (MAP-2, Sigma M4403) for 2hours in the same solution at the dilution of 1/400. This antibody wasrevealed with Alexa Fluor 488 goat anti-mouse (Molecular probe, ref: A11001) for 1 hour in the same solution.

Nuclei of cells were labelled by a fluorescent marker (Hoechst solution,SIGMA, ref: B1155).

For each condition, 20 pictures per well were taken using InCellAnalyzer™ 2000 (GE Healthcare) with 20× magnification. All images weretaken under the same conditions.

Statistics

The data were expressed as mean±s.e.mean (of 6 or 12 data percondition). A global analysis of the data was performed using a one-wayanalysis of variance (ANOVA). *p<0.05; **p<0.01; ***p<0.005.

Results

β amyloid 1-42 injury applied at 10 μM for 24 h induced a large andsignificant decrease (˜40%, p<0.005) in MAP 2 positive cells (FIG. 8).

Application of reference compound BDNF (50 ng/ml) inhibited cell deathresulting from β amyloid 1-42 injury. These results validate the study.

Pridopidine reduced the toxic effect of β amyloid 1-42 in a dosedependent manner. The proportion of neuron survival was 87% of themedium control when incubated with pridopidine at 200 μg/ml (p<0.005).At 100, 50, 25 and 10 μg/ml pridopidine increased significantly neuronsurvival resulting from β amyloid 1-42 injury (79%, p<0.005; 77%p<0.005; 74% p<0.01 and 71%, p<0.05, respectively).

Conclusions

Pridopidine increased neuron survival resulting from β amyloid 1-42injury significantly and with a dose dependent manner.

Example 19: Pridopidine Selectively Occupies the S1R in Humans at LowDose

Receptor Occupancy (RO) of pridopidine to the S1R and D2R was evaluatedusing Positron Emission Tomography (PET) imaging in the human brain(healthy volunteers and HD patients). The radio ligand(S)-(−)-[18F]fluspidine was used to image S1Rs, and the radio ligand[18F]fallypride was used to image D2Rs. The results of the imaginganalysis show that at a plasma exposure correlating to the 45 mg biddose steady state (ss) exposure, (90 mg single dose), pridopidine fullyoccupies the S1R (>90%) (FIG. 9).

Example 20: Pridopidine's S1R/D2R Occupancy and Brain Network ActivityDemonstrates Correlation

Correlation between S1R/D2R occupancy and brain network activity wasevaluated using the functional resting-state MRI (rs-fMRI). Functionalconnectivity (FC) was calculated from rs-fMRI data sets. Pridopidine lowdose (correlating to ss plasma exposure at 45 mg bid dose), shown by PETto selectively occupy only the S1R (no D2R binding), led to an increasein the DMN connectivity (increased activity) in healthy volunteers andin HD patients, as demonstrated in FIGS. 10-12. FIG. 10 showspridopidine increases FC within the DMN in healthy volunteers (HVs).This visual inspection result is supported by the results of theSPM-based voxelwise group comparison: FIG. 11: SPM-based voxelwise groupcomparison (within the DMN) in HV before (pre) and after (post) 90 mgpridopidine reveals pridopidine increased FC in the insula. FIG. 12:SPM-based voxelwise group comparison (within the DMN) in HD patientsbefore (pre) and after (post) 90 mg pridopidine reveals pridopidineincreased FC in the parietal and temporal cortex as well as precuneusand insula.

Pridopidine showed an unexpected increase in DMN at low dose whichselectively occupies the S1R.

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What is claimed is:
 1. A method of improving cognitive function in ahuman subject comprising administering to the subject a compositioncomprising pridopidine or a pharmaceutically acceptable salt thereofwherein the cognitive function in need for improvement is associatedwith DMN dysfunction.
 2. The method of claim 1, wherein the cognitivefunction is selected from the group consisting of global cognitivefunctioning, sustained cognition, memory, language, executivefunctioning, and attention.
 3. The method of claim 1, wherein thecognitive function is memory.
 4. The method of claim 3, wherein memoryis short term memory, long term memory or working memory.
 5. The methodof claim 1, wherein the subject is afflicted with a cognitive deficit.6. The method of claim 1, wherein the subject is prone to or predisposedto have a cognitive deficit.
 7. The method of claim 5, wherein thecognitive deficit is a memory deficit.
 8. The method of claim 7, whereinthe memory deficit is a short-term memory deficit or memory loss.
 9. Themethod of claim 8, wherein the memory loss is caused by one or more ofage-related changes in memory, mild cognitive impairment, dementia ordepression.
 10. The method of claim 5, wherein the cognitive deficit iscaused by or associated with a disease or disorder.
 11. The method ofclaim 10, wherein the disease or disorder is selected from the groupconsisting of Huntington's disease (HD), epilepsy, Mild cognitiveimpairment (MCI), Parkinson's disease, Alzheimer's disease, dementiaassociated with Lewy Bodies and Creutzfeldt-Jacob disease.
 12. Themethod of claim 1, wherein the amount of pridopidine or apharmaceutically acceptable salt thereof is administered to the humansubject once daily, 2-4 times a day, less than once a day, 2-4 times perweek.
 13. The method of claim 1, wherein the amount of pridopidineadministered is between 1 mg/day-315 mg/day, 22.5 mg/day-315 mg/day or90 mg/day-315 mg/day.
 14. The method of claim 1, wherein the amount ofpridopidine is administered orally, parenterally, intravascularly,paracancerally, transmucosally, transdermally, intramuscularly,intranasally, intravenously, intradermally, subcutaneously,sublingually, intraperitoneally, intraventricularly, intracranially,intravaginally, by inhalation, rectally, or intratumorally.
 15. Themethod of claim 1, wherein the pharmaceutically acceptable salt ofpridopidine is pridopidine hydrochloride, hydrobromide, nitrate,perchlorate, phosphate, sulphate, formate, acetate, aconate, ascorbate,benzenesulphonate, benzoate, cinnamate, citrate, embonate, enantate,fumarate, glutamate, glycolate, lactate, maleate, malonate, mandelate,methanesulphonate, naphthalene-2-sulphonate, phthalate, salicylate,sorbate, stearate, succinate, tartrate or toluene-p-sulphonate salt. 16.The method of claim 1, further comprising the administration of apsychiatric drug, an antidepressant drug, anxiolytics, antipsychoticmedications or combination thereof.
 17. The method of claim 16, whereinthe antidepressant is selected from the group consisting of citalopram,fluoxetine, paroxeine, sertraline, and trazodone.
 18. The method ofclaim 16, wherein the anxiolytic is selected from the group consistingof lorazepam and oxazepam.
 19. The method of claim 16, wherein theantipsychotic medication is selected from the group consisting ofaripiprazole, clozapine, haloperidol, olanzapine, quetiapine,risperidone, and ziprasidone.
 20. A method of treating a human subjectafflicted with epilepsy, comprising administering to the subject apharmaceutical composition comprising an amount of pridopidine or apharmaceutically acceptable salt thereof.
 21. The method of claim 20,wherein the composition is administered to the human subject once daily,2-4 times a day, less than once a day, 2-4 times per week.
 22. Themethod of claim 20, wherein the amount of pridopidine administered isbetween 1 mg/day-315 mg/day, 22.5 mg/day-315 mg/day or 90 mg/day-315mg/day.
 23. The method of claim 20, wherein the composition isadministered orally, parenterally, intravascularly, paracancerally,transmucosally, transdermally, intramuscularly, intranasally,intravenously, intradermally, subcutaneously, sublingually,intraperitoneally, intraventricularly, intracranially, intravaginally,by inhalation, rectally, or intratumorally.
 24. The method of claim 20,wherein the pharmaceutically acceptable salt of pridopidine ispridopidine hydrochloride, hydrobromide, nitrate, perchlorate,phosphate, sulphate, formate, acetate, aconate, ascorbate,benzenesulphonate, benzoate, cinnamate, citrate, embonate, enantate,fumarate, glutamate, glycolate, lactate, maleate, malonate, mandelate,methanesulphonate, naphthalene-2-sulphonate, phthalate, salicylate,sorbate, stearate, succinate, tartrate or toluene-p-sulphonate salt.